Synergistic effect of UV-A and UV-C light is traced to UV-induced damage of the transfer RNA.

UV light emitting diodes (LEDs) are considered the new frontier of UV water disinfection. As UV technologies continue to evolve, so does the need to understand disinfection mechanisms to ensure that UV treatment continues to adequately protect public health. In this research, two Escherichia coli (E. coli) strains (the wild type K12 MG1655 and K12 SP11 (ThiI E342K)) were irradiated with UV-C at 268 nm both independently and after exposure to UV-A (365 nm). A synergistic effect was found on the viability of the wild type E. coli K12 strain when UV-A irradiation was applied prior to UV-C. Sublethal UV-A doses, which had a negligible effect on cell viability alone, enhanced UV-C inactivation by several orders of magnitude. This indicated a specific cellular response mechanism to UV-A irradiation, which was traced to direct photolysis of the transfer RNA (tRNA), which are critical links in the translation of messenger RNA to proteins. The wild type K12 strain MG1655, containing tRNAs with a thiolated uridine, directly absorbs the UV-A light, which leads to a reduction in protein synthesis, making them more susceptible to UV-C induced damage. However, the K12 strain SP11 (ThiI E342K), with a point mutation in the thiI gene that prevents a post-transcriptional modification of tRNA, experienced less inactivation upon subsequent irradiation by UV-C. The growth rate of cells, which was inhibited by sublethal UV-A doses, was not inhibited in this mutant strain with the modified tRNA. Time-lapse microscopy with microfluidics showed that sub-lethal UV-A caused a transient, reversible, growth arrest in E. coli. However, once the growth resumed, the cell division time resembled that of unirradiated cells. Damage induced by UV-A impaired the recovery of damage induced by UV-C. Depending on the UV-A dose applied, the synergistic effect remained even when there was a time delay of several hours between UV-A and UV-C exposures. The effect of sublethal UV-A was reversible over time; therefore, the synergistic effect was strongest when UV-C was applied immediately after UV-A. Combining UV-A and UV-C irradiation may serve as a practical tool to increase UV disinfection efficacy, which could potentially reduce costs while still adequately protecting public health.

Evaluation of disinfection efficacy of single UV-C, and UV-A followed by UV-C LED irradiation on Escherichia coli, B. spizizenii and MS2 bacteriophage, in water.

Ultraviolet light-emitting diodes (UV LEDs) have shown ability to inactivate microorganisms and viruses in water. The unique characteristic of the UV-LEDs' diversity in wavelengths ranging from UV-C, UV-B, and UV-A, allows for wavelengths to be combined in different manners for polychromatic irradiation. Previous studies reported no synergy from simultaneous or sequential UV-C and UV-B as well as UV-C or UV-B followed by UV-A irradiation. However, synergy was reported for UV-A followed by UV-C or UV-B irradiation on various microorganisms. Nevertheless, no clear ground has been reached on whether to adopt single UV-C wavelengths or UV-A followed by UV-C LED, irradiation on inactivation of microorganisms and viruses in water. Therefore, this work evaluates the disinfection efficacy of single UV-C as well as UV-A followed by UV-C LED irradiation on Escherichia coli, Bacillus spizizenii spores and MS2 bacteriophage in water. The UV-C wavelengths were represented by 267 and 278 nm UV LEDs, and UV-A by 368 nm UV LEDs. In this study, E. coli was highly susceptible to UV radiation followed by B. spizizenii spores, and lastly MS2. Repair following UV inactivation was only observed in E. coli. The synergistic effect found in both E. coli, and B. spizizenii spores was attributed to the different inactivation mechanisms of the UV-C and UV-A wavelengths. In both single UV-C, and UV-A followed by UV-C LED irradiations, single 267 nm UV-C LED showed higher inactivation efficacy. Meanwhile, single 278 nm UV-C LED showed higher efficacy in terms of suppression of repair, and electrical energy consumption. Using single UV-C LEDs in a water disinfection system cuts down on related extra costs by avoiding combined wavelengths while still attaining better levels of microorganism inactivation, repair suppression and electrical energy consumption. These findings are applicable for the design and implementation of UV LED water disinfection systems.

Efficacy of UVC-LED in water disinfection on Bacillus species with consideration of antibiotic resistance issue.

Ultraviolet light emitting diode (UV-LED) has attracted extensive attention as a new technology to replace traditional mercury lamp for water disinfection. This study reported for the first time the application of UVC-LEDs in range of 200-280 nm for the treatment of two Gram-positive tetracycline resistant bacteria (TRB) from Bacillus species and their tetracycline resistant gene (TRG). The results showed that UVC-LEDs can inactivate TRB up to 5.7-log and inhibit TRG expression, especially at 268 nm. The required fluence was approximate to that of the referential non-resistant bacteria using the same UVC-LED, but far less than that of TRB using mercury lamp. After UVC-LED irradiation, photoreactivation was the dominant mechanism to repair TRB, just like non-resistant bacteria. But contrary to non-resistant bacteria, the regrowth ratio of TRB was remarkably high at 24 h since the end of the irradition, nevertheless the number of the regrown bacteria in the irradiated water was still less than that in the non-irradiated water. Whereas TRB restored resistance after repair even applying 268 nm at a fluence up to 46.08 mJ/cm2 (maximum in this study). This study highlights the merits of UVC-LED to effectively inactivate TRB in a prompt, energy-efficient and resistance-reducing way, while future study on TRB regrowth and resistance resilience is needed.

Comparison of the performance of pulsed and continuous UVC-LED irradiation in the inactivation of bacteria.

Ultraviolet light-emitting diode (UV-LED) is a newly emerging UV light source with a potential to replace the conventional chemical methods, mercury UV lamps and xenon lamps in water disinfection. UV-LEDs are characterized by the diversity in wavelengths and can be turned on and off with a high and adjustable frequency, making them an attractive candidate for pulsed light (PL) disinfection apart from the conventional continuous wave (CW) operation. Previous studies on comparison between the PL and CW UV-LED irradiations for the inactivation of bacterium in water disinfection are limited and results conflict. In this work, PL and CW UV-LED in the C-spectral band (UVC-LED) irradiations were compared at equivalent fluence in the inactivation and subsequent photoreactivation of E. coli bacteria. In addition, effect of different driving currents and ambient temperatures on solder temperature of the PL and CW UVC-LED irradiation was also examined. Under the equivalent fluences, the PL and CW UVC-LED irradiations brought comparable inactivation efficiency and similar photoreactivation of the E. coli. Moreover, the PL UVC-LED exhibited a much lower solder temperature than the CW UVC-LED irradiation. On the other hand, CW UVC-LED irradiation, higher ambient temperature and higher driving current increased the solder temperature that resulted to a negative impact on the wavelength, full width at half maximum, optical power and irradiance, which are key factors in the UVC-LED disinfection efficiency. In all, this work reports the comparison between PL and CW in UVC-LED irradiation for inactivating E. coli bacteria and firstly revealed the photoreactivation of the E. coli after the PL UVC-LEDs irradiation.

Effects of single and combined UV-LEDs on inactivation and subsequent reactivation of E. coli in water disinfection.

Ultraviolet light emitting diodes (UV-LEDs) have shown a potential to replace traditional Ultraviolet (UV) pressure lamps for water disinfection. However, the research is not sufficient and hence, it is still difficult to make any logical conclusions. In this work, UV-LEDs with peak emissions at 267, 275, 310 nm and combined emissions at 267/275, 267/310 and 275/310 nm were applied to a batch water disinfection system. Under either single- or combined-wavelength situation, the inactivation efficiency, reactivation (due to photoreactivation and dark repair) after UV irradiation and electrical energy consumption were evaluated by way of the model bacterium Escherichia coli. It was found that, the 267 nm UV-LED had the highest inactivation efficiency than other UV-LEDs. Although reactivation occurred after 267, 275, 267/275 and 275/310 nm UV-LEDs' irradiations, it occurred to a lesser extent in dark repair than in photoreactivation, demonstrating that photo-effect is the dominant mechanism of reactivation. In addition, decay phase was more prominent than reactivation in dark repair. However, the irradiation by the 275 nm UV-LED showed a better persistence against reactivation which could be attributed to protein damage at 275 nm. No synergistic effect for combined wavelengths was observed in this study. The electrical energy consumption was lower for the 275 nm UV-LED than the other UV-LEDs which was attributed to its higher wall plug efficiency. This study showed the variation principle between the single and combined UVB/UVC-LEDs in inactivation efficiency, inhibition of reactivation, synergistic effect and electrical energy consumption in treatment of E. coli, which is useful for the reasonable exploitation of UV-LEDs in water disinfection systems.